Antifouling peptides combined with recognizing DNA probes for ultralow fouling electrochemical detection of cancer biomarkers in human bodily fluids.

Herein, a universal strategy for the construction of highly sensitive and low fouling biosensors was proposed based on antifouling peptides conjugated with recognizing DNA probes. The peptide-DNA conjugate was formed through a reagent-free click reaction between a typical DNA aptamer modified with 5'-dibenzocyclooctyne (DBCO) and the designed antifouling peptide terminated with biotin and the azide group at its two ends. With the assistance of streptavidin (SA), the electrochemical biosensor was constructed via immobilization of the straight peptides and peptide-DNA conjugates in sequence onto the electrode surface modified with electrodeposited poly(3,4-ethylenedioxythiophene) (PEDOT) and gold nanoparticles (AuNPs). The prepared biosensor exhibited excellent antifouling performances in various human bodily fluids such as serum, sweat and urine, with a wide linear response range for CA125 from 0.01 U mL to 1000 U mL, and a low limit of detection of 0.003 U mL. Combining the advantages of the antifouling peptide and recognizing DNA probe, this sensing strategy was capable of assaying CA125 in undiluted human serum, and it also offered a highly promising way for the development of different antifouling biosensors through the conjugation of antifouling peptides with various DNA probes.